3 results about "Cell dissociation" patented technology

The application discloses a kind of high efficiency gentle mammal blastocyst single cell dissociation method.The method includes the following steps: (1) zona pellucida removal: the blastocyst is placed in calcium magnesium table type liquid, and zona pellucida is completely removed by dissolving and physical blowing operation;(2) enzyme digestion incubation: the blastocyst after step (1) is handled and is transferred to 35-37 ℃ pre-equilibrated complex dissociation solution and incubated for 15-30 minutes, during which intermittent blowing is carried out, until the blastocyst is completely dissociated into single cell;(3) washing: the single cell obtained by dissociation is washed using dPBS, and the residual dissociation solution is removed.The method of the application has high efficiency, high activity and simplified operation.The dissociation process is simplified from 3-4 steps to two steps (zona pellucida dissolution+enzyme digestion incubation), the total time is shortened from more than 90 minutes to less than 30 minutes, the dissociation efficiency is significantly improved;Cell viability is increased from about 70% to more than 90%, the cell junction is gently lysed, and cell damage is reduced.

The present application relates to the field of biotechnology, in particular to human bladder tissue digestion solution and application thereof.The digestion method of the present application can dissociate human bladder tissue cells while keeping the cell viability at more than 90%, and the amount of harvested cells is also obviously improved.Sufficient cell amount is the prerequisite for optimization treatment such as red blood cell lysis and dead cell removal.The single cell suspension obtained by the method has high viability, low clumping rate, high nucleated rate and less impurities, which can provide a basis for obtaining high-quality gene expression data in human bladder tissue single cell sequencing experiment.The present application can quickly obtain human bladder tissue primary cell suspension (sufficient cell amount, high viability, low clumping rate and less impurities) meeting the requirements of single cell experiment within 2-4 hours.

This invention provides a kit, method, and single-cell library construction method for the dissociation of marine animal tissues. The dissociation method includes: S1) dissociating marine animal tissue samples at 8℃~12℃ using a cell dissociation buffer to obtain a single-cell suspension; S2) centrifuging the single-cell suspension and washing the single-cell precipitate obtained after centrifugation with a cell resuspension buffer to obtain single cells from the marine animal tissue sample; wherein, the cell dissociation buffer comprises a protease; the protease is Bacillus subtilis protease A; the concentration of sodium ions in the cell resuspension buffer is 300-800mM, and the concentration of potassium ions is 5-20mM.